Background:Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease.Theref...Background:Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease.Therefore,development of a sensitive technique for the diagnosis of schistosomiasis in domestic animals is necessary.Method:A novel colloidal gold immunochromatography assay(GICA)strip was developed for detecting Schistosoma japonicum in domestic animals.The colloidal gold was conjugated with recombinant streptococcal protein G(rSPG).As the test and control lines,the schistosome soluble egg antigen and rSPG,respectively,were blotted on nitrocellulose membrane.Results:The lowest detectable serum dilution was 1∶640 for schistosome-infected buffaloes.The cross-reaction rate of GICA was 14.29%with Paramphistomum sp.in buffaloes,16.67%with Haemonchus sp.in goats,and 33.33%with Orientobilharzia sp.in goats.These results were slightly lower and similar to those obtained through ELISA.Moreover,the strips for detecting S.japonicum in mice,rabbits,buffaloes,and goats showed high sensitivity(100.00%,100.00%,100.00%,and 100.00%,respectively)and specificity(100.00%,100.00%,94.23%,and 88.64%,respectively).And the sensitivity or specificity of the GICA strips did not present any significant differences after storage for 12 months at room temperature.When compared with ELISA,the GICA strips exhibited similar sensitivity and specificity in the diagnosis of schistosomiasis in mice,rabbits,buffaloes,and goats.Besides,only 5μl of serum are required for the test and the detection can be completed within 5 min.Conclusion:This study is the first to develop a GICA strip using gold-rSPG conjugate for the diagnosing of schistosomiasis in domestic animals,and preliminary results showed that the developed strip may be suitable for large-scale screening of schistosomiasis in endemic areas.展开更多
pH plays a vital role in various cellular activities, and real-time observation of the intracellular p H through a p H indicator is very important for studying many physiological processes. In this paper, we studied t...pH plays a vital role in various cellular activities, and real-time observation of the intracellular p H through a p H indicator is very important for studying many physiological processes. In this paper, we studied the p H response of Trp–Trp dipeptide and its derivatives(NATrp_2Me, NBTrp_2 and Trp_2Me) by steady-state and time-resolved fluorescence spectroscopy. Both the fluorescence intensities and lifetimes of Trp–Trp dipeptide as well as Trp2 Me were functions of p H in the physiological range from 5.5 to 9.0. However, NATrp_2 Me and NBTrp_2 showed no difference. The exposed amino was found to be pivotal for its p H dependence. Moreover, an artificially synthesized tetrapeptide(Trp–Trp–Ala–Ser) confirmed the p H sensitivity of N-terminal Trp–Trp residues. The p H values could be quantitatively determined from the fluorescence intensities and lifetimes of the N-terminal Trp–Trp residue. Thus, the N-terminal Trp–Trp residues may be fused into the polypeptides/proteins to serve as an intrinsic p H indicator in fluorescence spectroscopy and imaging.展开更多
基金This study was financially supported by the Basic Foundation for Scientific Research of State-level Public Welfare Institutes of China(Grant no.2014JB01).
文摘Background:Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease.Therefore,development of a sensitive technique for the diagnosis of schistosomiasis in domestic animals is necessary.Method:A novel colloidal gold immunochromatography assay(GICA)strip was developed for detecting Schistosoma japonicum in domestic animals.The colloidal gold was conjugated with recombinant streptococcal protein G(rSPG).As the test and control lines,the schistosome soluble egg antigen and rSPG,respectively,were blotted on nitrocellulose membrane.Results:The lowest detectable serum dilution was 1∶640 for schistosome-infected buffaloes.The cross-reaction rate of GICA was 14.29%with Paramphistomum sp.in buffaloes,16.67%with Haemonchus sp.in goats,and 33.33%with Orientobilharzia sp.in goats.These results were slightly lower and similar to those obtained through ELISA.Moreover,the strips for detecting S.japonicum in mice,rabbits,buffaloes,and goats showed high sensitivity(100.00%,100.00%,100.00%,and 100.00%,respectively)and specificity(100.00%,100.00%,94.23%,and 88.64%,respectively).And the sensitivity or specificity of the GICA strips did not present any significant differences after storage for 12 months at room temperature.When compared with ELISA,the GICA strips exhibited similar sensitivity and specificity in the diagnosis of schistosomiasis in mice,rabbits,buffaloes,and goats.Besides,only 5μl of serum are required for the test and the detection can be completed within 5 min.Conclusion:This study is the first to develop a GICA strip using gold-rSPG conjugate for the diagnosing of schistosomiasis in domestic animals,and preliminary results showed that the developed strip may be suitable for large-scale screening of schistosomiasis in endemic areas.
基金supported by the National Natural Science Foundation of China(6110807761178085)+2 种基金the Science and Technology Commission of Shanghai Municipality(1552071150015ZR1411700)the Program of Introducing Talents of Discipline to Universities(B12024)
文摘pH plays a vital role in various cellular activities, and real-time observation of the intracellular p H through a p H indicator is very important for studying many physiological processes. In this paper, we studied the p H response of Trp–Trp dipeptide and its derivatives(NATrp_2Me, NBTrp_2 and Trp_2Me) by steady-state and time-resolved fluorescence spectroscopy. Both the fluorescence intensities and lifetimes of Trp–Trp dipeptide as well as Trp2 Me were functions of p H in the physiological range from 5.5 to 9.0. However, NATrp_2 Me and NBTrp_2 showed no difference. The exposed amino was found to be pivotal for its p H dependence. Moreover, an artificially synthesized tetrapeptide(Trp–Trp–Ala–Ser) confirmed the p H sensitivity of N-terminal Trp–Trp residues. The p H values could be quantitatively determined from the fluorescence intensities and lifetimes of the N-terminal Trp–Trp residue. Thus, the N-terminal Trp–Trp residues may be fused into the polypeptides/proteins to serve as an intrinsic p H indicator in fluorescence spectroscopy and imaging.
